Document Type: Original Article

Authors

1 Laboratoire de Génie Microbiologique et Applications, Université des frères Mentouri, Constantine, Algérie

2 Laboratoire Bioqual, Institut de la Nutrition, de l’Alimentation et des Technologies Agro-Alimentaires, Université des frères Mentouri, Constantine, Algérie

3 Unité des Rickettsies, Faculté de Médecine, Université de la Méditerranée, Marseille, France

10.26655/ijabbr.2016.9.7

Abstract

Actinomycetes are an uncommon agent of human infections and its pathogenic factors are not known. The present study reports a rare case isolation of an actinomycete from a woman pleural fluid; the strain was identified by 16S rRNA gene sequence analysis. This strain was tested to produce an extracellular protease that hydrolysis gelatin, casein and hemoglobin on agar mediums. The purification of the enzyme was carried by ammonium sulfate precipitation, gel filtration and ion exchange chromatographies. The activity of protease was studied at different pH values and temperatures and in the presence of metallic ions and inhibitors. The molecular weight of the enzyme was determined by 12% Tricine SDS-polyacrylamide gel electrophoresis. The strain was identified as Streptomyces cinereoruber ssp. cinereoruber. Extracellular proteolytic enzyme was purified at 19.67 fold and a 3.0% recovery. The enzyme was characterized as having optimal activities at pH 11.0 and 50°C, it keeps more than 50% of activity at pH between 4.0 to 12.0 and it is thermostable at 30 and 40°C. Enzymatic activity is enhanced in the presence of metal ions and inhibited by EDTA and 1,10-phenanthroline. The molecular weight was 53 kDa. This study reports the first case isolation of Streptomyces cinereoruber ssp. cinereoruber from pleural fluid, the extracellular zinc-metalloprotease was proposed as candidate virulence factor.

Keywords

Main Subjects

Armstrong, P.B., 2006. Proteases and protease inhibitors: a balance of activities in host–pathogen interaction. Immunobiol., 211, 263–281.

Cheng, G., He, L., Sun, Z., Cui, Z., Du, Y., Kong, Y., 2015. Purification and biochemical characterization of a novel fibrinolytic enzyme from Streptomyces sp. P3. J. Microbiol. Biotechnol., 25, 1449–1459.

Chitte, R., Dey, S., 2000. Potent fibrinolytic enzyme from a thermophilic Streptomyces megasporus strain SD5. Lett. Appl. Microbiol., 31, 405-410.

Dunne, E.F., Burman, W.J., Wilson, M.L., 1998. Streptomyces pneumonia in patient with human immunodeficiency virus infection: Case report and review of the literature on invasive Streptomyces infections. Clin. Infec. Dis., 27, 93–96.

El-Shanshoury, A.E.R.R., El-Sayed, M.A., Sammour, R.H., El-Shouny, W.A., 1995. Purification and partial characterization of two extracellular alkaline proteases from Streptomyces corchorusii ST36. Can. J. Microbiol., 41, 99-104.

Felsenstein, J., 1985. Confidence limits on phylogenies: an approach using the bootstrap. Evol., 39, 783-791.

Gagaoua, M., Boucherba, N., Bouanane-Darenfed, A., Ziane, F., Nait-Rabah, S., Hafid, K., Boudechicha, H.R., 2014. Three-phase partitioning as an efficient method for the purification and recovery of ficin from mediterranean fig (Ficus carica L.) latex. Separ. Purif. Technol., 132, 461–467.

Gugnani, A.C., Unaogu, I.C., Emeruwa, C.N., 1993. Pulmonary infection due to Streptomyces griseus. J. Commun. Dis., 25, 38–40.

Hui, Z., Doi, H., Kanouchi, H., Matsuura, Y., Mohri, S., Nonomura, Y., Oka, T., 2004. Alkaline serine protease produced by Streptomyces sp. degrades PrPSc. Bioch. Bioph. Res. Commun., 321, 45-50.

Kang, S.G., Kim, I.S., Rho, Y.T., Lee, K.J., 1995. Production dynamics of extracellular proteases accompanying morphological differentiation of Streptomyces albidoflavus SMF301. Microbiol., 141, 3095-3103.

Kapadia, M., Rolston, K., Han, X.Y., 2007. Invasive Streptomyces infections: six cases and literature review. Am. J. Clin. Pathol., 127, 619–624.

Kato, J.Y., Suzuki, A., Yamazaki, H., Ohnishi, Y., Horinouchi, S., 2002. Control by A-factor of a metalloendopeptidase gene involved in aerial mycelium formation in Streptomyces griseus. J. Bacteriol., 184, 6016-6025.

Lee, S., Kim, K., 2012. Protease activity: meeting its theranostic potential. Theranostics, 2, 125-126.

Lowry, O.H., Rosebrough, N.J., Farr, A.L., Randall, R.J., 1951. Protein measurement with the folin-phenol reagents. J. Biol. Chem., 193, 265-275.

Manteca, A., Pelaeza, A.I., Garcia-Suareza, M.D.M., Hidalgob, E., Bustob, B.D., Mendeza, F.J., 2008. A rare case of lung coinfection by Streptomyces cinereoruber and Haemophilus influenzae in a patient with severe chronic obstructive pulmonary disease: Characterization at species level using molecular techniques. Diag. Microbiol. Infec. Dis., 60, 307–311.

Martin, G.J., Blazes, D.L., Mayers, D.L., Spooner, K.M., 1999.  Refractory craniofacial actinomycetoma due to Streptomyces somaliensis that required salvage therapy with amikacin and imipenem. Clin. Infec. Dis., 29, 460–461.

McNeil, M.M., Brown, J.M., 1994. The medically important aerobic actinomycetes: Epidemiology and microbiology. Clin. Microbiol. Rev., 7, 357-417.

Moreira, K.A., Cavalcanti, M.T.H., Duarte, H.S., Tambourgi, E.B., Magalhães de Melo, E.H., Silva, V.L., Porto, A.L.F., Filho, J.L.D.L., 2001. Partial characterization of proteases from Streptomyces clavuligerus using an inexpensive medium. Braz. J. Microbiol., 32, 215-220.

Pellegrini, G.J., Graziano, J.C., Ragunathan, L., Bhat, M.A., Hemashettar, B.M.,  Browna, J.M., 2012. Scalp abscess due to Streptomyces cacaoi subsp. cacaoi, first report in a human infection. J. Clin. Microbiol., 50, 1484-1486.

Ramesh, S., Rajesh, M., Mathivanan, N., 2009. Characterization of a thermostable alkaline protease produced by marine Streptomyces fungicidicus MML1614. Biopross. Biosys. Eng., 32, 791-800.

Riviere, E., Neau, D., Roux, X., Lippa, N., Roger-Schmeltz, J., Mercie, P., Longy Boursier, M., 2012. Pulmonary Streptomyces infection in patient with sarcoidosis, France, Emerg. Infect. Dis., 18, 1907-1909.

Rose III, C.E., Brown, J.M., Fisher, J.F., 2008. Brain abscess caused by Streptomyces infection following penetration trauma: Case report and results of susceptibility analysis of 92 isolates of Streptomyces species submitted to the CDC from 2000 to 2004. J. Clin. Microbiol., 46, 821–823.

Saitou, N., Nei, M., 1987. The neighbor-joining method-a new method for reconstructing phylogenetic trees. Mol. Biol. Evol., 4, 406-425.